LOCUS       sequence-330645-        6362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Puromycin resistance and LifeAct-mCherry expression for Salpingoeca 
            rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  AUTHORS   Brunet T, Albert M, Roman W, Coyle MC, Spitzer DC, King N
  TITLE     A flagellate-to-amoeboid switch in the closest living relatives of 
            animals.
  JOURNAL   Elife. 2021 Jan 15;10. pii: 61037. doi: 10.7554/eLife.61037.
  PUBMED    33448265
REFERENCE   2  (bases 1 to 6362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2021-01-15"; volume: "10. pii: 61037. doi"; pages: " 
            10.7554/eLife.61037"
COMMENT     Sequence Label: NK803
FEATURES             Location/Qualifiers
     source          1..6362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..51
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     primer_bind     complement(1426..1446)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3072..3089)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3072..3088)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3081..3103)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3297..3316)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3416..3438
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3476..3494)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3562..3666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3667..3735,3736..4527)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3885..3904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4698..5286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5187..5206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5440..5457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5574..5595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5610..5615,5616..5633,5634..5640)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5648..5664
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5653..5675
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5672..5688
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5672..5688
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgggtgttg ctgaccttat taagaaattc gagtctatct ccaaggaaga gggcggtagc
       61 ggtggatcta tggtctccaa gggcgaggag gacaacatgg ctatcatcaa ggagttcatg
      121 cgcttcaagg tccacatgga gggctccgtc aacggccacg agttcgagat cgagggcgag
      181 ggcgagggcc gcccctacga gggcacccag accgccaagc tcaaggtgac gaagggcggc
      241 cccctcccct tcgcctggga catcctctcc ccccagttca tgtacggctc caaggcttac
      301 gtcaagcacc ccgccgacat ccccgactac ctcaagctct ccttccccga gggcttcaag
      361 tgggagcgcg tcatgaactt cgaggacggc ggcgtcgtca ccgtcaccca ggactcctct
      421 cttcaggacg gcgagttcat ctacaaggtc aagctgcgcg gcaccaactt cccctccgac
      481 ggccccgtca tgcagaagaa gacgatgggt tgggaggctt cctccgagcg catgtacccc
      541 gaggacggcg ccctcaaggg cgagatcaag cagcgcctca agctcaagga cggcggccac
      601 tacgacgccg aggtcaagac cacctacaag gccaagaagc ccgtccagct ccccggcgcc
      661 tacaacgtca acatcaagct cgacatcacc tcccacaacg aggactacac tatcgttgag
      721 cagtatgagc gtgccgaggg tcgtcacagc acgggcggta tggacgagct ttacaagtct
      781 taagagctcc ccccagcatt atcacgtcta ctgtgtgtat gttgtcgctc tctgtttgac
      841 ctctctcttg tctactatgt tgcctctctc tctttcttga ttgcacccct ttcctcgcta
      901 cgactactac gacttccgct accttgactt tgttggcttg ttgccacctt ttggtcttgc
      961 gtttcgtctg ctcgtcactc ttgtctcttt ttgacgactt gtgtgtgatg acggcgtgag
     1021 gctgtgactg actgactctc aacaacttcc aataaaaacc cctacatcgc acgtgactga
     1081 ctgcacattt agtgaaggga aggaaaagaa gtgaggtgtt ttcatgtaca tgaggcttgt
     1141 cgttgctcgg gcaccgctca ttgatcgtca cagttataca ttggggggaa ggatagttac
     1201 actggcatag tgggtttatt gcggggccca agggctcggc atgacgtgca caagaagcag
     1261 aatcaactca caacaagcgc acgtggcacg cgtcgacacg aggacaacag caaaagaggc
     1321 aacaaggtgg caatgaccaa cgaggcgcac cgggcttgcg ggtcatgcac caggtacgag
     1381 gaccctcggg gacctccacg tcggcggtga cggtgaagcc gagacgctcg tagaagggga
     1441 ggttgcgagg agcggaggtc tcgaggaaag cagggacacc agcacgctca gcggcctcga
     1501 cgccagggag gacgacggca gagccgagac ccttgccctg gtggtcagga gagacgccga
     1561 cggtggcgag gaaccaggcg ggctccttgg ggcggtgggg ggcgaggagg ccctccatct
     1621 gctgctgagc ggcgagacgg gagccagaca gctcagccat gcgggggccg atctcagcga
     1681 agacagcacc ggcctcgaca gactcagggg tggtccagac agcgacagca gcaccgtcgt
     1741 cggcgaccca gaccttcccg atgtcgaggc cgacgcgggt gaggaacagc tcctgcagct
     1801 cggtgacacg ctcgatgtgg cggtcagggt cgacggtgtg acgggtagca gggtagtcag
     1861 cgaaggcagc ggcgagggta cggacagcac ggggcacgtc gtcacgggta gcgaggcgga
     1921 cggtaggctt gtactccgta ccagacatgt tgctggttgt tgatggtgtt gacggtaaac
     1981 ggggtcaaag aagaagacga cgcgaaaaga ctgctcacgt ggcacgagga aaaagagcag
     2041 agaaataaat ggtgttggtg gacggagaag gggaggaagg aaggaaggca ggaaggaagt
     2101 caggagggga agaaaaaaag gcaaggaaag cgacgagaag ggaaaggcat ttgtgagcgg
     2161 acaatgtgaa cgtttcacgt gcgtgtgtat tgtgtgtgcg tgcgtgcgtg tgtgcttgag
     2221 cgctggcttg tattggtgat ggcttgtatt ggttatgttg gtggtgtgaa acataacaca
     2281 cgaccatgcg tgcgagcaat gcttttgtgc gggtgcggtg gtggctgtct tttatctcgt
     2341 cgtcattgct cgcccttccc ttcccttcac ctctttccta gtctcctctc ctctccctca
     2401 tccttgtcgc tcttcgcgtg gctcgccttg caccaaccac caaccaccaa ccatacaacc
     2461 accaccacct ctcatcatcc atagaagctc actcatatcc gtcgacgaaa ccaacctctt
     2521 gagctgcaga ctgactcaat gaaaaaggca gatgcacatg cgccaatgcc gcaaacccct
     2581 ctccttgttt ctcgcagcca ctctgcgtgt aacaggctga gtgagcagaa gcaagaacga
     2641 agccacacgc ggccacgcca cgcacacagg ccagacacaa gcccagcaag cacttttttt
     2701 ttcatgggga ttgtgcggga gtttgtgttg tcgacagcga gcacgtggtc gcatctttcg
     2761 caacattcgc gtttctgggc cagaaacggg tccagaaacg ggtcgtgtgg tacgggccca
     2821 agttgtgtcg gcaggaaatt tggtgcaaat cacacacaaa tcagcgtgcg cccatgacaa
     2881 agaggcgcgc aagcaatgca cgcgaacctt cctcgccatt tgtgctgact cgacacgttg
     2941 gtatctgttg caaattgtac agaagtcact gttgcaattg gcaaaagttg ggtgggagac
     3001 agagaacgac gacgacaaac gacaacaaga ctgcagtcgt tacaggatcc ccgggtaccg
     3061 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3121 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3181 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     3241 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     3301 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     3361 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3421 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     3481 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3541 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3601 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3661 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3721 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3781 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3841 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3901 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3961 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4021 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4081 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4141 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4201 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4261 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4321 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4381 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4441 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4501 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4561 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4621 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4681 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4741 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4801 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4861 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4921 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4981 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5041 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5101 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5161 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5221 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5281 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5341 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5401 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5461 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5521 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     5581 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     5641 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     5701 aagcttgtaa caaactgccc tgcacctcgt ctgccacctc ggttgccacc ttgtcctcgt
     5761 tccccaccac cttgtcctcg tccgctgcca gcatcgcgat cagcgtgctt gcgcttggcg
     5821 ctgctgctgc cggacttccc gttatcggcc atactttttg tggtgtttcc aggcgagatg
     5881 cgctgttggc aaagcaaaat ggcaaaacgg cctttgccac acgccatcct cgcctcacct
     5941 cgacgtgcgg ctttcctaaa caggaaacct gacgaaacga attgagcgca ttggcacacg
     6001 tgtgctgtgc gctttgtgtc gctgcatgct atccaccacg caccacccaa tcagtccaac
     6061 tgtgacgtgt cacgcctagg cgaaaaagcg gcgagagcgc caaagcaaac gccaaacaaa
     6121 aaccaaagaa gagcaaacgc agcgaactga tgtgagacaa gggggcggtg caatgcgtca
     6181 ctgaacgtgc aaaagccaaa aacaaccaag aagcagctgc acaacacaac aacaacaaca
     6241 acaacaacaa caccttgttg tcgcagctct acgtttggca ttttcctgtt gttctctctt
     6301 gcgatcagtc cgtgtctctt ttctccccct tttcccagac cacaaccaca aaacaaccag
     6361 cc
//
